CN112120712B - Preparation kit for Platelet Rich Plasma (PRP) of leucocyte - Google Patents

Preparation kit for Platelet Rich Plasma (PRP) of leucocyte Download PDF

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Publication number
CN112120712B
CN112120712B CN202011014055.XA CN202011014055A CN112120712B CN 112120712 B CN112120712 B CN 112120712B CN 202011014055 A CN202011014055 A CN 202011014055A CN 112120712 B CN112120712 B CN 112120712B
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platelet
rich plasma
centrifuge tube
centrifugal machine
centrifuge
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CN112120712A (en
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敖英芳
闫文强
胡晓青
龚熹
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Peking University Third Hospital Peking University Third Clinical Medical College
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Peking University Third Hospital Peking University Third Clinical Medical College
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/15Devices for taking samples of blood
    • A61B5/150007Details
    • A61B5/150755Blood sample preparation for further analysis, e.g. by separating blood components or by mixing
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/15Devices for taking samples of blood
    • A61B5/150007Details
    • A61B5/150015Source of blood
    • A61B5/15003Source of blood for venous or arterial blood
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/15Devices for taking samples of blood
    • A61B5/153Devices specially adapted for taking samples of venous or arterial blood, e.g. with syringes
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention provides a preparation set of Platelet Rich Plasma (PRP) with white blood cells, which comprises a rubber tourniquet, a first injector, a blood taking needle, a second injector, an anticoagulant storage device, a platelet rich plasma extraction centrifuge tube and a centrifuge; the centrifuge tube comprises: the centrifugal tube comprises a centrifugal tube body with a centrifugal tube cavity, a platelet leukocyte removal filter membrane arranged in the centrifugal tube cavity, a three-way tube arranged on the centrifugal tube body and positioned below the leukocyte removal filter membrane, and a centrifugal tube cover detachably connected to a tube orifice of the centrifugal tube body; the tee includes: an upper longitudinal duct and a lower longitudinal duct, and a transverse duct in communication with the upper longitudinal duct; one end of the serum transfer pipeline extends into the centrifuge tube cavity for a certain length, the other end of the serum transfer pipeline is connected with the port of the lower longitudinal pipeline, the upper end of the upper longitudinal pipeline is communicated with the lower end of the centrifuge tube cavity, and the three-way pipe is connected with a valve. The PRP extracted by the application is a leucocyte PRP, and has no organism inflammatory response caused by pro-inflammatory factors generated by leucocytes.

Description

Preparation kit for Platelet Rich Plasma (PRP) of leucocyte
Technical Field
The invention relates to the technical field of medical devices, in particular to a preparation set of Platelet Rich Plasma (PRP) with white blood cells.
Background
Platelet rich plasma (platelet rich plasma, PRP) is a platelet concentrate obtained from whole blood by centrifugation, contains high concentration of platelets, and can release a large amount of growth factors after platelet activation to promote body healing, so that the platelet rich plasma has been widely used clinically, for example, sports injury (arthritis, synovitis, tennis elbow, etc.), primary surgery (open wound healing, intra-traumatic fixation, etc.), secondary surgery (delayed fracture healing, bone nonunion, etc.). Currently, the PRP extraction kit which is commonly used and already produced contains a large amount of white blood cells because white blood cells are not removed, and the literature shows that proinflammatory factors secreted by cells can trigger inflammatory reactions of organisms, thereby influencing healing of the organisms.
Disclosure of Invention
The invention provides a preparation kit of Platelet Rich Plasma (PRP) of leucocytes, which is used for solving the technical problems proposed by the background technology.
In order to solve the technical problems, the invention discloses a preparation set of Platelet Rich Plasma (PRP) with white blood cells, which comprises a rubber tourniquet, a blood taking needle, a first injector, a second injector, an anticoagulant storage device, a platelet rich plasma extraction centrifuge tube and a centrifuge, wherein the blood taking needle is detachably connected with the first injector;
The platelet rich plasma extraction centrifuge tube comprises: the centrifugal tube comprises a centrifugal tube body with a centrifugal tube cavity, a platelet leukocyte removal filter membrane arranged in the centrifugal tube cavity, a three-way tube arranged on the centrifugal tube body and positioned below the leukocyte removal filter membrane, and a centrifugal tube cover detachably connected to a tube orifice of the centrifugal tube body;
the tee includes: an upper longitudinal duct and a lower longitudinal duct, and a transverse duct connected between the upper longitudinal duct and the lower longitudinal duct and communicating with the upper longitudinal duct; the blood serum transfer pipeline, one end of blood serum transfer pipeline stretches into the centrifuging tube intracavity a certain length, the other end of blood serum transfer pipeline with the port connection of lower longitudinal pipeline, the upper end and the centrifuging tube chamber lower extreme intercommunication of going up longitudinal pipeline, the tee bend pipe is connected with the valve.
Preferably, the centrifuge comprises:
a host;
the rotating shaft is rotatably arranged in the host machine and is driven by a driving motor in the host machine, and a plurality of rotors are rotatably arranged in the host machine;
the first connecting rods are in one-to-one correspondence with the rotors, one end of each first connecting rod is fixedly connected with the corresponding rotating shaft, the other end of each first connecting rod is provided with a fixed ring, and the rotors are rotationally connected to the inner sides of the fixed rings through connecting shafts arranged in front of and behind the first connecting rods;
The outer centrifuge tubes are provided with rotor centrifuge cavities, and the rotor centrifuge cavities are provided with the platelet-rich plasma extraction centrifuge tubes;
and the rotor centrifugal cavity cover is detachably connected with the outer centrifugal tube and is used for closing or opening the rotor centrifugal cavity.
Preferably, the first syringe is a 20ml syringe for collecting whole blood; the second injector is a 10ml injector and is used for extracting bottom red blood cells, surface serum and PRP;
the anticoagulant stored in the anticoagulant storage device is 4% sodium citrate anticoagulant.
Preferably, the centrifuge tube cover is provided with a first connecting tube, an external thread is arranged at the outer part of the upper end of the first connecting tube, and a screw cap is connected at the external thread in a threaded manner and is used for sealing the first connecting tube.
Preferably, the leukocyte-removing filter membrane is a concentrated platelet leukocyte-removing polyester filter membrane.
Preferably, the method for using the leukocyte-depleted platelet-rich plasma (PRP) preparation kit comprises the following steps:
step 1: obtaining whole blood, comprising:
step 11: 2ml of 4% sodium citrate anticoagulant was withdrawn using the 20ml syringe;
step 12: the rubber tourniquet is tightly tied above the elbow joint, after the vein of the elbow is filled, the blood taking needle is used for venipuncture, the tail part of the blood taking needle is connected with a 20ml syringe after the step 11 is completed, and 18ml of whole blood is extracted;
Step 13: unscrewing a screw cap above the platelet and leukocyte removing filter membrane, connecting a 20ml syringe in the first connecting pipe to filter the whole blood by the platelet and leukocyte removing filter membrane, and tilting the centrifuge tube for 30 degrees to enable the whole blood to flow to the bottom of the tube along the tube wall of the platelet-rich plasma extraction centrifuge tube;
step 2: blood centrifugation stratification, comprising:
placing the platelet-rich plasma extraction centrifuge tube completed in the step 1 into a centrifuge cavity of a rotor of a centrifuge, screwing a rotor centrifuge cavity cover, and adjusting centrifuge parameters to be: 1000g, 10min, 4 ℃; after centrifugation, the blood is layered, and comprises red blood cells at the bottom layer, platelet layers in the middle and serum at the uppermost layer;
step 3: obtaining platelet rich plasma, comprising:
step 31: rotating a three-way pipe valve, opening a transverse pipeline of the three-way pipe, using the 10ml syringe to extract bottom red blood cells to the 2ml scale of the centrifuge tube, namely reserving 2ml of bottom red blood cells, and closing the transverse pipeline;
step 32: then rotating the valve, opening the lower longitudinal pipeline of the three-way pipe, and extracting upper serum through the serum transfer pipeline by using the 10ml syringe;
step 33: the lateral tubing was again opened and concentrated platelet rich plasma was withdrawn using a 10ml syringe.
Preferably, the device further comprises a cleaning device for cleaning the platelet-rich plasma extraction centrifuge tube, wherein the platelet-rich plasma extraction centrifuge tube is clamped with a platelet-removing leukocyte filter membrane, and the cleaning device comprises:
the upper end of the mounting seat is provided with a plurality of mounting cavities for mounting the platelet-rich plasma extraction centrifuge tube;
the fixed bracket is fixedly connected to one side of the upper end of the mounting seat, and the upper end of the fixed bracket is fixedly connected with a horizontal fixed plate;
the left fixing block and the right fixing block are fixedly connected to the lower end of the horizontal fixing plate at intervals;
the threaded rod is horizontally arranged, two ends of the threaded rod are rotationally connected with the two fixed blocks, a first thread and a second thread are respectively arranged on the outer walls of the left side and the right side of the center of the threaded rod, the rotation directions of the first thread and the second thread are opposite, and the threaded rod is driven by a first rotating motor arranged on the fixed support or the horizontal fixed plate;
the two sliding blocks are provided with horizontal threaded through holes, and the horizontal threaded through holes of the two sliding blocks are respectively connected to the first threads and the second threads in a threaded manner;
the second mounting plate is arranged below the sliding block;
The upper ends of the two second connecting rods are respectively connected with the two sliding blocks in a rotating way, and the lower ends of the two second connecting rods are respectively connected to the second mounting plate in a rotating way;
the upper ends of the first vertical fixing rods are connected to the second mounting plate, liquid inlet cavities are formed in the first vertical fixing rods, and cleaning and clamping assemblies are arranged on the first vertical fixing rods;
the cleaning and clamping assembly includes:
the arc plates are arranged at intervals on the periphery of the lower end of the vertical fixing rod, the upper ends of the arc plates are rotationally connected with the arc plates, and the lower ends of the arc plates are provided with hook parts;
the brush hair is arranged on the outer side of the arc-shaped plate, a connecting block is fixedly connected to the inner side of the arc-shaped plate, and the inner side surface of the connecting block is a first conical surface;
the outer side of the pushing block is provided with a second conical surface, the outer side of the pushing block is abutted with the first conical surface, and the lower end of the pushing block is provided with a pushing ball;
the second fixing rod is internally provided with a vertical flow passage, and the vertical flow passage is communicated with the liquid inlet cavity, and the upper end of the second fixing rod is fixedly connected to the lower end of the first vertical fixing rod;
the spring is sleeved outside the second fixing rod, the upper end of the spring is fixedly connected with the lower end of the first vertical fixing rod, and the upper end of the spring is fixedly connected with the upper end of the pushing block;
The miniature water pump is arranged on the mounting seat or the fixing support or the horizontal fixing plate, the water inlet end of the miniature water pump is connected with a water source, the water outlet end of the miniature water pump is connected with the liquid inlet cavity through a second connecting pipe, and an electromagnetic valve is arranged on the second connecting pipe;
the displacement sensor is arranged on the horizontal fixing plate and used for detecting the vertical displacement of the second mounting plate;
and the microcontroller is arranged on the mounting seat or the fixed support or the horizontal fixed plate and is electrically connected with the miniature water pump, the displacement sensor and the first rotating motor.
Preferably, the upper end of the first vertical fixing rod is rotatably connected to the second mounting plate, a second rotating motor for driving the first vertical fixing rod to rotate is arranged on the second mounting plate, and the second connecting pipe is arranged at the center of the upper end of the first vertical fixing rod.
Preferably, the method further comprises:
the pressure sensors are respectively arranged at the front side, the upper side and the left side of the contact position of the connecting shaft and the rotor;
a first detecting device for detecting the rotation speed of the driving motor;
the second detection device is used for detecting the flow rate of the whole blood in the platelet-rich plasma extraction centrifuge tube;
The third detection device is used for detecting an included angle between the axis of the rotor and the horizontal direction in the longitudinal rotation process of the rotor;
the controller is electrically connected with the pressure sensor, the first detection device, the second detection device and the third detection device, and the controller controls the driving motor to work with the controller based on the pressure sensor, the first detection device, the second detection device and the third detection device, and comprises the following steps:
step 1: the controller calculates the separation efficiency of the centrifugal machine based on the second detection device, the third detection device and the formula (1);
Figure GDA0004179426260000051
wherein K is the separation efficiency of the centrifuge (7), e is a natural constant, the value is 2.71828, ρ is the whole blood density, D is the diameter of a filter hole of a platelet leukocyte removal filter membrane, D is the diameter of a platelet-rich plasma extraction centrifuge tube, μ is the dynamic viscosity of whole blood, sin is the sine alpha is the maximum included angle between the axis of the rotor and the horizontal direction in the longitudinal rotation process of the rotor, v is the flow velocity of the whole blood in the platelet-rich plasma extraction centrifuge tube, η is the ratio of the whole blood volume in the platelet-rich plasma extraction centrifuge tube to the total volume in the platelet-rich plasma extraction centrifuge tube, A 1 For the current rotation speed of the driving motor of the centrifugal machine, A 3 The rated rotation speed of the driving motor of the centrifugal machine is set;
step 2: the controller calculates a safety evaluation coefficient Q of the centrifugal machine based on the step 1, the third detection device, the pressure sensor and the formula (2);
Figure GDA0004179426260000061
wherein A is 2 For the maximum safe rotation speed of the driving motor of the centrifugal machine, x is the average value of all the pressure sensor detection values at the front side, y is the average value of all the pressure sensor detection values at the upper side, z is the average value of all the pressure sensor detection values at the left side, and max (x, y, z) is the maximum value of the three values of x, y and z;
step 3: the controller compares the separation efficiency of the centrifugal machine with a preset separation efficiency reference value, and compares the safety evaluation coefficient of the centrifugal machine with a preset safety evaluation coefficient reference value;
when the separation efficiency of the centrifugal machine is greater than or equal to a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is greater than or equal to a preset safety evaluation coefficient reference value, the controller does not trigger an alarm device;
when the separation efficiency of the centrifugal machine is smaller than a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is smaller than a preset safety evaluation coefficient reference value, the controller controls the alarm device to send out a first alarm prompt;
When the separation efficiency of the centrifugal machine is greater than or equal to a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is smaller than the preset safety evaluation coefficient reference value, the controller controls the alarm device to send out a second alarm prompt, and the rotating speed of the driving motor of the centrifugal machine is reduced;
when the separation efficiency of the centrifugal machine is smaller than a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is larger than or equal to the preset safety evaluation coefficient reference value, the controller controls to increase the rotating speed of the centrifugal machine, and when the rotating speed is increased so that the separation efficiency of the centrifugal machine is equal to the preset separation efficiency reference value, the controller calculates the safety evaluation coefficient of the centrifugal machine again, if the safety evaluation coefficient of the centrifugal machine is larger than or equal to the preset safety evaluation coefficient reference value at the moment, the controller does not trigger an alarm device, and if the safety evaluation coefficient of the centrifugal machine is smaller than the preset safety evaluation coefficient reference value at the moment, the controller controls the alarm device to send a third alarm prompt.
The technical scheme of the invention is further described in detail through the drawings and the embodiments.
Drawings
The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate the invention and together with the embodiments of the invention, serve to explain the invention. In the drawings:
FIG. 1 is a schematic diagram of a part of the device assembly structure before the nested centrifugation of the present invention.
FIG. 2 is a schematic diagram of a part of the device assembly during the centrifugation of the kit according to the present invention.
Fig. 3 is a schematic structural view of the connection of the rotor of the centrifuge with the rotating shaft, the connecting shaft and the fixing ring according to the present invention.
Fig. 4 is a schematic diagram showing a change of a movement state of a rotor of the centrifugal machine connected with a rotating shaft.
Fig. 5 is a schematic structural view of an embodiment of the cleaning device of the present invention.
FIG. 6 is a schematic view of a portion of the cleaning and clamping assembly of FIG. 5.
In the figure: 1. a rubber tourniquet; 2. a blood taking needle; 3. a first syringe; 4. a second syringe; 5. an anticoagulant storage device; 6. platelet rich plasma extraction centrifuge tube; 61. a centrifuge tube body; 62. a centrifuge tube cover; 63. a first connecting pipe; 7. a centrifuge; 71. a host; 72. a rotating shaft; 73. a first connecting rod; 74. a fixing ring; 75. a connecting shaft; 76. an outer centrifuge tube; 77. a rotor centrifugal chamber cover; 78. a rotor; 8. a three-way pipe; 81. an upper longitudinal duct; 82. a lower longitudinal duct; 83. a transverse duct; 9. a serum transfer line; 10. a cleaning device; 101. a mounting base; 102. a mounting cavity; 103. a fixed bracket; 104. a horizontal fixing plate; 105. a fixed block; 106. a threaded rod; 107. a slide block; 108. a second mounting plate; 109. a second connecting rod; 110. a first vertical fixing rod; 111. cleaning and clamping the assembly; 1111. an arc-shaped plate; 1112. a connecting block; 1113. a pushing block; 1114. pushing the ball; 1115. a second vertical fixing rod; 1116. and (3) a spring.
Detailed Description
The preferred embodiments of the present invention will be described below with reference to the accompanying drawings, it being understood that the preferred embodiments described herein are for illustration and explanation of the present invention only, and are not intended to limit the present invention.
In addition, the descriptions of the "first," "second," and the like, herein are for descriptive purposes only and are not intended to be specifically construed as order or sequence, nor are they intended to limit the invention solely for distinguishing between components or operations described in the same technical term, but are not to be construed as indicating or implying any relative importance or order of such features. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include at least one such feature. In addition, technical solutions and technical features between the embodiments may be combined with each other, but it is necessary to base that a person skilled in the art can implement the combination of technical solutions, when the combination of technical solutions contradicts or cannot be implemented, should be considered that the combination of technical solutions does not exist, and is not within the scope of protection claimed by the present invention.
Example 1
The invention discloses a preparation set of Platelet Rich Plasma (PRP) of leucocyte, as shown in figures 1-3,
The blood taking device comprises a rubber tourniquet 1, a blood taking needle 2, a first injector 3, a second injector 4, an anticoagulant storage device 5, a platelet-rich plasma extraction centrifuge tube 6 and a centrifuge 7, wherein the blood taking needle 2 is detachably connected with the first injector 3; alternatively, the lancet may be a butterfly needle;
the platelet rich plasma extraction centrifuge tube 6 comprises: a centrifuge tube body 61 having a centrifuge tube cavity (for accommodating blood), a platelet leukocyte-removing filter membrane mounted in the centrifuge tube cavity, a tee 8 mounted on the centrifuge tube body 61 and located below the leukocyte-removing filter membrane (specifically, as shown in fig. 1, at the lower end of the centrifuge tube body), a centrifuge tube cap 62 detachably attached to a tube orifice of the centrifuge tube body 61 (for sealing the tube orifice);
the tee 8 includes: an upper longitudinal duct 81 and a lower longitudinal duct 82, and a transverse duct 83 connected between the upper longitudinal duct 81 and the lower longitudinal duct 82 and communicating with the upper longitudinal duct; the serum transfer pipeline 9, the one end of serum transfer pipeline 9 stretches into the centrifuging tube intracavity certain length, the other end of serum transfer pipeline 9 with the port connection of lower longitudinal pipe 82, the upper end and the centrifuging tube intracavity lower extreme intercommunication of upper longitudinal pipe 81, tee pipe 8 is connected with the valve. Preferably, valves may be respectively provided at the discharge nozzles (ports) of the lower longitudinal pipes and the discharge nozzles (ports) of the lateral pipes;
The rubber tourniquet comprises: bundling the elbow to fill the vein of the elbow, thereby facilitating venipuncture; the blood taking needle is used for venipuncture and drawing blood in a patient; the first injector 3 is used for collecting whole blood; the second injector 4 is used for extracting bottom red blood cells, surface serum and PRP;
wherein the first syringe 3 may be, but is not limited to, a 20ml syringe, and the second syringe 4 may be, but is not limited to, a 10ml syringe;
the anticoagulant stored in the anticoagulant storage means 5 may be, but is not limited to, a 4% sodium citrate anticoagulant for in vitro anticoagulation of raw plasma;
the concentrated platelet leukocyte-removing filter membrane can be, but is not limited to, a concentrated platelet leukocyte-removing polyester filter membrane;
the concentrated platelet leukocyte-removing filter membrane described above is used to filter out leukocytes in whole blood, and to retain platelets.
Preferably, a first connecting pipe 63 is arranged on the centrifugal pipe cover 62, an external thread is arranged outside the upper end of the first connecting pipe 63, and a nut is connected at the external thread in a threaded manner and is used for sealing the first connecting pipe 63; the first connecting pipe is arranged, so that when whole blood is injected into the centrifuge tube body through the second injector, only the screw cap is required to be opened, the whole centrifuge tube cover is not required to be opened, and the tightness is improved; and a connecting pipe is arranged, so that the second injector is more convenient to be connected with one position of the connecting pipe for injecting the ring blood.
The working principle and beneficial effects of the technical scheme are as follows: the invention is used for filtering white blood cells in whole blood by arranging the concentrated platelet leukocyte-removing filter membrane and retaining platelets. Solves the problems that the prior PRP extraction set which is commonly used and has been produced has a large number of white blood cells because of no clearance of white blood cells, and the existing literature shows that the proinflammatory factors secreted by the cells can trigger inflammatory reaction of organisms, thereby influencing the healing of the organisms.
The blood is centrifuged and then sequentially from top to bottom: serum (platelet-poor plasma layer), platelet layer (buffy coat layer), erythrocytes; providing the tee pipe comprising an upper longitudinal pipe 81 and a lower longitudinal pipe 82, and a transverse pipe 83 connected between the upper longitudinal pipe 81 and the lower longitudinal pipe 82 and communicating with the upper longitudinal pipe; a serum transfer pipeline 9, wherein one end of the serum transfer pipeline 9 extends into the centrifuge tube cavity for a certain length, the other end of the serum transfer pipeline 9 is connected with a port of the lower longitudinal pipeline 82, the upper end of the upper longitudinal pipeline 81 is communicated with the lower end of the centrifuge tube cavity, and the three-way pipe 8 is connected with a valve; the blood serum transfer pipeline is a runner in the three-way pipe, and the blood serum is only discharged through a discharge pipe orifice at the lower end of the lower longitudinal pipeline through the blood serum transfer pipeline; the upper end of the upper longitudinal pipe is communicated with the lower end of the centrifuge tube cavity, and the upper longitudinal pipe is communicated with (only) the transverse pipe, so that another flow channel is formed, and the platelet layer and the red blood cells can be discharged from the other flow channel; the above technical scheme is convenient for separating serum, platelet layer (buffy coat layer) and red blood cells.
The technology contained in the application can effectively filter white blood cells before centrifugation, and the platelets are retained to the greatest extent.
In summary, the present application has the following advantages: 1. the PRP extracted by the application is derived from the blood of a patient and is used by the patient, so that the risk of immune rejection and disease transmission is avoided;
2. the PRP extracted by the application is a leucocyte PRP, and has no organism inflammatory reaction caused by pro-inflammatory factors generated by leucocytes;
3. according to the method, the PRP containing the high-concentration platelets is obtained by removing redundant red blood cells and serum, so that a large amount of growth factors can be generated, and the healing of organisms is promoted;
4. the PRP extracted by the patent can be combined with thrombin or calcium chloride to form PRP gel, and the PRP gel is adhered to a tissue defect part to promote local formation of a high-concentration growth factor environment, and provide a support for cell migration, so that the repair of the tissue defect is facilitated.
5. The equipment that this application relates to is simple in manufacturing to all adopt green material preparation, obviously reduce manufacturing cost.
Example 2
On the basis of example 1, as shown in FIGS. 2-3
The centrifuge 7 includes:
a host 71;
the rotating shaft 72 is rotatably arranged in the host machine and driven by a driving motor in the host machine, and the host machine 71 is rotatably provided with a plurality of rotors 78;
The first connecting rods 73 are in one-to-one correspondence with the rotors 78, one end of each first connecting rod 73 is fixedly connected with the corresponding rotating shaft 72, a fixed ring 74 is arranged at the other end of each first connecting rod 73, and the rotors 78 are rotatably connected to the inner side of the fixed ring 74 through connecting shafts 75 arranged front and back;
the outer centrifuge tubes 76 are provided with rotor 78 centrifuge cavities, and the rotor 78 centrifuge cavities are provided with the platelet rich plasma extraction centrifuge tubes;
and a rotor centrifugal cavity cover 77 detachably connected with the outer centrifugal tube and used for closing or opening the rotor 78 centrifugal cavity.
The working principle and beneficial effects of the technical scheme are as follows: in the initial state, the outer centrifuge tube is vertically arranged as shown in fig. 4; the driving motor drives the rotating shaft to horizontally rotate, so that an integrated structure which is connected with the rotating shaft and comprises a first connecting rod, a rotor, an outer centrifuge tube and a fixing ring starts to horizontally rotate; simultaneously, in the rotating process, the rotor longitudinally rotates around the connecting shaft due to centrifugal force, and the rotating speed of the driving motor can be set, so that the rotor is positioned at the horizontal position shown in fig. 4; the combination of the horizontal rotation and the longitudinal rotation can accelerate the whole blood stratification;
at present, most PRP extraction sets used at home and abroad adopt secondary centrifugation operation to extract PRP, and secondary centrifugation obviously prolongs the operation time and the in-vitro residence time of the PRP, thereby influencing the activity of growth factors in the PRP. According to the method, the PRP can be obtained by accelerating whole blood stratification and adopting a single centrifugation technology, so that the operation time can be obviously shortened, the in-vitro residence time of the PRP is reduced, and the activity of the growth factors in the PRP is reserved to the greatest extent.
The rotor centrifugal cavity cover is arranged for sealing the rotor centrifugal cavity, so that the safety coefficient during centrifugation can be improved;
example 3
Based on example 1 or 2;
the using method of the leucocyte-rich platelet (PRP) preparation kit comprises the following steps:
step 1: obtaining whole blood, comprising:
step 11: 2ml of 4% sodium citrate anticoagulant was withdrawn using the 20ml syringe;
step 12: the rubber tourniquet 1 is fastened above the elbow joint of a patient, after the elbow vein is filled, the blood taking needle 2 is used for venipuncture, the tail part of the blood taking needle 2 is connected with a 20ml syringe after the step 11 is completed, and 18ml of whole blood is extracted;
step 13: unscrewing a screw cap above the platelet and leukocyte removing filter membrane, connecting a 20ml syringe in the first connecting pipe 63, filtering whole blood by the platelet and leukocyte removing filter membrane, and simultaneously tilting the centrifuge tube for 30 degrees to enable the whole blood to flow to the bottom of the tube along the tube wall of the platelet-rich plasma extraction centrifuge tube;
step 2: blood centrifugation stratification, comprising:
placing the platelet rich plasma extraction centrifuge tube finished in the step 1 into a centrifugal cavity (in particular a centrifugal cavity of an outer centrifuge tube) of a rotor 78 of a centrifuge 7, screwing a rotor centrifugal cavity cover, and adjusting centrifugal parameters to be: 1000g, 10min, 4 ℃; after centrifugation, the blood is layered, and comprises red blood cells at the bottom layer, platelet layers in the middle and serum at the uppermost layer;
Step 3: to obtain a concentrated platelet-rich plasma, it is necessary to remove a part of the bottom red blood cells and the surface serum in order to increase the platelet concentration. Comprising the following steps:
step 31: rotating the valve of the three-way pipe 8, opening the transverse pipeline 83 of the three-way pipe 8, using the 10ml syringe to extract the bottom red blood cells to the 2ml scale of the centrifuge tube, namely reserving 2ml of bottom red blood cells, and closing the transverse pipeline 83;
step 32: subsequently, the valve is rotated, the lower longitudinal pipe 82 of the tee pipe 8 is opened, and the 10ml syringe is used for extracting the upper serum through the serum transfer pipe 9;
step 33: the lateral channel 83 was again opened and concentrated platelet rich plasma was withdrawn using a 10ml syringe.
The working principle and beneficial effects of the technical scheme are as follows:
the whole operation flow is simple, and the damage to the patient is small;
the blood serum transfer pipeline is a runner in the three-way pipe, and the blood serum is only discharged through a discharge pipe orifice at the lower end of the lower longitudinal pipeline through the blood serum transfer pipeline; the upper end of the upper longitudinal pipe is communicated with the lower end of the centrifuge tube cavity, and the upper longitudinal pipe is communicated with (only) the transverse pipe, so that another flow channel is formed, and the platelet layer and the red blood cells can be discharged from the other flow channel; the technical proposal is convenient for separating serum, platelet layer (buffy coat) and red blood cells. The technical scheme is convenient for separating and operating, and is convenient for separating serum, platelet layer (buffy coat layer) and red blood cells.
Example 4
On the basis of any one of embodiments 1 to 3, as shown in fig. 5 to 6, the apparatus further comprises a cleaning device 10, configured to clean the platelet-rich plasma extraction centrifuge tube 6, wherein a platelet-removing leukocyte-removing filter membrane is clamped on a sidewall of the platelet-rich plasma extraction centrifuge tube 6 (specifically, the sidewall of the centrifuge tube may have a structure in which a clamping groove is provided, a block matched with the clamping groove is provided on the filter membrane, and the clamping block may be an elastic clamping block), where the cleaning device 10 includes:
the mounting seat 101 is provided with a plurality of mounting cavities 102 at the upper end of the mounting seat 101 for mounting the platelet-rich plasma extraction centrifuge tube 6;
the fixed support 103 is fixedly connected to one side of the upper end of the mounting seat 101, and the upper end of the fixed support 103 is fixedly connected with a horizontal fixed plate 104;
the two fixing blocks 105 are fixedly connected to the lower end of the horizontal fixing plate 104 at left and right horizontal intervals;
the threaded rod 106 is horizontally arranged, two ends of the threaded rod 106 are rotatably connected with the two fixing blocks 105, a first thread and a second thread are respectively arranged on the outer walls of the threaded rod 106, which are positioned on the left side and the right side of the center of the threaded rod 106, the rotation directions of the first thread and the second thread are opposite, and the threaded rod 106 is driven by a first rotating motor arranged on the fixing support 103 or the horizontal fixing plate 104;
The two sliding blocks 107 are provided with horizontal threaded through holes, and the horizontal threaded through holes of the two sliding blocks 107 are respectively connected to the first threads and the second threads in a threaded manner;
a second mounting plate 108 disposed below the slider 107;
the upper ends of the two second connecting rods 109 are respectively and rotatably connected with the two sliding blocks 107, and the lower ends of the two second connecting rods 109 are respectively and rotatably connected with the second mounting plate 108;
the upper ends of the first vertical fixing rods 110 are connected to the second mounting plate 108, liquid inlet cavities are formed in the first vertical fixing rods 110, and cleaning and clamping assemblies 111 are arranged on the first vertical fixing rods 110;
the cleaning and clamping assembly 111 includes:
the arc plates 1111 are arranged at intervals on the periphery of the lower end of the vertical fixing rod, the upper ends of the arc plates 1111 are rotatably connected with the arc plates 1111, and the lower ends of the arc plates 1111 are provided with hook parts for hooking the filter membrane;
bristles are arranged on the outer side of the arc-shaped plate 1111, a connecting block 1112 is fixedly connected to the inner side of the arc-shaped plate 1111, and the inner side surface of the connecting block 1112 is a first conical surface;
the outer side of the pushing block 1113 is a second conical surface, the outer side of the pushing block 1113 is abutted against the first conical surface, and a pushing ball 1114 is arranged at the lower end of the pushing block 1113;
The second fixing rod 1115 is internally provided with a vertical flow channel, and the vertical flow channel is communicated with the liquid inlet cavity, and the upper end of the second fixing rod is fixedly connected to the lower end of the first vertical fixing rod 110;
the spring 1116 is sleeved outside the second fixed rod, the upper end of the spring 1116 is fixedly connected with the lower end of the first vertical fixed rod 110, and the upper end of the spring 1116 is fixedly connected with the upper end of the pushing block 1113;
the miniature water pump is arranged on the mounting seat 101 or the fixing support or the horizontal fixing plate 104, the water inlet end of the miniature water pump is connected with a water source, the water outlet end of the miniature water pump is connected with the liquid inlet cavity through a second connecting pipe (preferably, a connecting hose comprising a hard pipe and one end of the hard pipe, the hard pipe is convenient for arranging a valve, the hose is convenient for the movement of a first vertical fixing rod), and the second connecting pipe is provided with an electromagnetic valve;
a displacement sensor disposed on the horizontal fixing plate 104 for detecting a vertical displacement of the second mounting plate;
and the microcontroller is arranged on the mounting seat 101 or the fixed support or the horizontal fixed plate 104 and is electrically connected with the miniature water pump, the displacement sensor and the first rotating motor.
The working principle and beneficial effects of the technical scheme are as follows:
the hook part can be arranged outside, when the platelet-rich plasma extraction centrifuge tube 6 is used and needs to be cleaned, the platelet-rich plasma extraction centrifuge tube 6 is placed in the mounting cavity, and a plurality of platelet-rich plasma extraction centrifuge tubes 6 can be placed in the mounting cavities at the same time so as to accelerate the cleaning efficiency; after the placing is finished, a user starts a microcontroller, the microcontroller controls the first rotating motor to rotate positively, two sliding blocks on the threaded rod are close to each other, so that the second mounting plate and the structure on the second mounting plate are lowered, at the moment, the vertical displacement of the second mounting plate is sensed by the displacement sensor, when the second mounting plate is lowered, the cleaning and clamping assembly is driven to descend into the platelet-rich plasma extraction centrifuge tube 6, at the moment, the lower ends of the arc plates in the cleaning and clamping assembly are close to each other, the cleaning and clamping assembly is convenient to enter the centrifuge tube, when the hook parts at the lower ends of the arc plates of the cleaning and clamping assembly reach the platelet-rich plasma extraction filter membrane, a miniature water pump is started at the moment, the cleaning liquid is pumped, so that the cleaning liquid enters the pushing block from the upper part, the pushing ball on the pushing block is pushed by the gravity of the cleaning liquid, the connecting plate and the arc plates are pushed to the outside, so that the hook parts at the inner sides of the lower ends of the arc plates rotate outwards, and the platelet-rich plasma extraction filter membrane at the moment, and the platelet-rich plasma extraction centrifuge tube is detached from the platelet-rich plasma extraction centrifuge tube 6; simultaneously, the brush hair on the outer side of the arc-shaped plate is in close contact with the inner side wall of the platelet-rich plasma extraction centrifuge tube 6, and the cleaning liquid can wet the brush hair, so that the inner wall of the platelet-rich plasma extraction centrifuge tube 6 can be cleaned more conveniently; at the moment, the microcontroller controls the first rotating motor to rotate reversely at intervals, so that the arc-shaped plate and bristles on the arc-shaped plate move upwards to clean the inner wall of the centrifugal tube, and meanwhile, the filter membrane is driven to move upwards, and the filter membrane is taken out from the centrifugal tube; after the blood plasma extraction centrifugal tube is taken out, the microcontroller controls the first rotating motor to positively turn over at intervals, and meanwhile, cleaning liquid is continuously input into the centrifugal tube through the water pump, so that the arc-shaped plate and bristles on the arc-shaped plate move up and down to clean the inner wall of the platelet-rich plasma extraction centrifugal tube 6; the technical scheme realizes automatic cleaning of the centrifuge tube and automatic removal of the filter membrane, and is convenient for repeated use of the centrifuge tube.
Example 5
On the basis of any one of examples 1 to 4,
the upper end of the first vertical fixing rod 110 is rotatably connected to the second mounting plate 108, a second rotating motor for driving the first vertical fixing rod 110 to rotate is arranged on the second mounting plate 108, and the second connecting pipe is arranged at the center of the upper end of the first vertical fixing rod 110.
The working principle and beneficial effects of the technical scheme are as follows: the upper end of the first vertical fixing rod 110 is rotatably connected to the second mounting plate 108, and a second rotating motor for driving the first vertical fixing rod 110 to rotate is arranged on the second mounting plate 108, so that the horizontal rotation cleaning is convenient while the up-and-down movement cleaning is realized; the second connecting pipe is arranged at the center of the upper end of the first vertical fixing rod 110, so that liquid is fed in the horizontal rotation process.
Example 6
On the basis of any one of embodiments 1 to 5, further comprising:
the pressure sensors are respectively arranged at the front side, the upper side and the left side of the contact part of the connecting shaft (75) and the rotor (78);
a first detecting device for detecting the rotation speed of the driving motor;
a second detecting device for detecting the flow rate of whole blood in the platelet rich plasma extraction centrifuge tube 6;
The third detecting device is configured to detect an included angle between the axis of the rotor 78 and the horizontal direction during the longitudinal rotation of the rotor 78;
the controller is electrically connected with the pressure sensor, the first detection device, the second detection device and the third detection device, and the controller controls the driving motor to work with the controller based on the pressure sensor, the first detection device, the second detection device and the third detection device, and comprises the following steps:
step 1: the controller calculates the separation efficiency of the centrifuge 7 based on the second detection device, the third detection device and the formula (1);
Figure GDA0004179426260000161
wherein K is the separation efficiency of the centrifuge (7), e is a natural constant, the value is 2.71828, ρ is the whole blood density, D is the diameter of the filter hole of the platelet-removing leukocyte filter membrane, D is the diameter of the platelet-rich plasma extraction centrifuge tube 6, μ is the dynamic viscosity of whole blood, sin is the sine alpha is the maximum included angle between the axis of the rotor and the horizontal direction in the longitudinal rotation process of the rotor, v is the flow velocity of the whole blood in the platelet-rich plasma extraction centrifuge tube, and η is the flow velocity of the whole blood in the platelet-rich plasma extraction centrifuge tubeRatio of Whole blood volume to total volume in platelet rich plasma extraction centrifuge tube, A 1 For the current rotation speed of the driving motor of the centrifugal machine, A 3 The rated rotation speed of the driving motor of the centrifugal machine is set;
step 2: the controller calculates a safety evaluation coefficient Q of the centrifugal machine based on the step 1, the third detection device, the pressure sensor and the formula (2);
Figure GDA0004179426260000162
wherein A is 2 For the maximum safe rotation speed of the driving motor of the centrifugal machine, x is the average value of all the pressure sensor detection values at the front side, y is the average value of all the pressure sensor detection values at the upper side, z is the average value of all the pressure sensor detection values at the left side, and max (x, y, z) is the maximum value of the three values of x, y and z;
step 3: the controller compares the separation efficiency of the centrifugal machine with a preset separation efficiency reference value, and compares the safety evaluation coefficient of the centrifugal machine with a preset safety evaluation coefficient reference value;
when the separation efficiency of the centrifugal machine is greater than or equal to a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is greater than or equal to a preset safety evaluation coefficient reference value, the controller does not trigger an alarm device;
when the separation efficiency of the centrifugal machine is smaller than a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is smaller than a preset safety evaluation coefficient reference value, the controller controls the alarm device to send out a first alarm prompt;
When the separation efficiency of the centrifugal machine is greater than or equal to a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is smaller than the preset safety evaluation coefficient reference value, the controller controls the alarm device to send out a second alarm prompt, and the rotating speed of the driving motor of the centrifugal machine is reduced;
when the separation efficiency of the centrifugal machine is smaller than a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is larger than or equal to the preset safety evaluation coefficient reference value, the controller controls to increase the rotating speed of the centrifugal machine, and when the rotating speed is increased so that the separation efficiency of the centrifugal machine is equal to the preset separation efficiency reference value, the controller calculates the safety evaluation coefficient of the centrifugal machine again, if the safety evaluation coefficient of the centrifugal machine is larger than or equal to the preset safety evaluation coefficient reference value at the moment, the controller does not trigger an alarm device, and if the safety evaluation coefficient of the centrifugal machine is smaller than the preset safety evaluation coefficient reference value at the moment, the controller controls the alarm device to send a third alarm prompt.
The working principle and beneficial effects of the technical scheme are as follows: detecting the rotating speed of the driving motor through a first detection device; detecting the flow rate of whole blood in the platelet rich plasma extraction centrifuge tube by a second detection device; detecting an included angle between the axis of the rotor and the horizontal direction in the longitudinal rotation process of the rotor through a third detection device; the controller controls the driving motor and the controller to work based on the pressure sensor, the first detection device, the second detection device and the third detection device;
First,: the controller calculates the separation efficiency of the centrifugal machine based on the second detection device, the third detection device and the formula (1); and the comprehensive consideration in the formula (1): the method comprises the steps of calculating separation efficiency by the influences of the whole blood density, the diameter of a filter hole of a platelet-rich plasma extraction centrifuge tube, the diameter of the filter hole of the platelet-rich plasma extraction centrifuge tube, the dynamic viscosity of whole blood, the maximum included angle between a rotor axis and the horizontal direction in the longitudinal rotation process of a rotor, the flow velocity of the whole blood in the platelet-rich plasma extraction centrifuge tube, the ratio of the whole blood volume in the platelet-rich plasma extraction centrifuge tube to the total volume in the platelet-rich plasma extraction centrifuge tube, the current rotation speed of a driving motor of the centrifuge and the rated rotation speed of the driving motor of the centrifuge, so that the separation efficiency is matched with the real-time specific whole blood factors, the rotation motor rotation speed factors of the centrifuge and the longitudinal rotation factors of the rotor, and the calculated separation efficiency is more accurate;
then the controller calculates a safety evaluation coefficient Q of the centrifugal machine based on the step 1, the third detection device, the pressure sensor and the formula (2), and the formula (2) comprehensively considers the comparison of the separation efficiency, the maximum safety rotation speed of the driving motor of the centrifugal machine and the actual rotation speed; and the actual stress factors of the front side, the upper side and the left side of the contact position of the connecting shaft and the rotor 78 to calculate the safety evaluation coefficient, so that the stress and the separation efficiency are considered and the safety rotation speed is considered during the safety evaluation, and the safety evaluation is more reliable;
Finally, the controller compares the separation efficiency of the centrifugal machine with a preset separation efficiency reference value, and compares the safety evaluation coefficient of the centrifugal machine with a preset safety evaluation coefficient reference value;
when the separation efficiency of the centrifugal machine is greater than or equal to a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is greater than or equal to a preset safety evaluation coefficient reference value, the controller does not trigger an alarm device; when the separation efficiency of the centrifugal machine is smaller than a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is smaller than a preset safety evaluation coefficient reference value, the controller controls the alarm device to send out a first alarm prompt; when the separation efficiency of the centrifugal machine is greater than or equal to a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is smaller than the preset safety evaluation coefficient reference value, the controller controls the alarm device to send out a second alarm prompt, and the rotating speed of the driving motor of the centrifugal machine is reduced; when the separation efficiency of the centrifugal machine is smaller than a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is larger than or equal to the preset safety evaluation coefficient reference value, the controller controls to increase the rotating speed of the centrifugal machine, and when the rotating speed is increased so that the separation efficiency of the centrifugal machine is equal to the preset separation efficiency reference value, the controller calculates the safety evaluation coefficient of the centrifugal machine again, if the safety evaluation coefficient of the centrifugal machine is larger than or equal to the preset safety evaluation coefficient reference value at the moment, the controller does not trigger an alarm device, and if the safety evaluation coefficient of the centrifugal machine is smaller than the preset safety evaluation coefficient reference value at the moment, the controller controls the alarm device to send a third alarm prompt. Different working conditions are prompted in real time through the comparison, management staff can take different countermeasures to deal with in time through different prompts, centrifugal efficiency is guaranteed when the centrifugal machine works, meanwhile, the working safety of the centrifugal machine is not affected, and the centrifugal machine is safer to use.
It will be apparent to those skilled in the art that various modifications and variations can be made to the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present invention also include such modifications and alterations insofar as they come within the scope of the appended claims or the equivalents thereof.

Claims (7)

1. A preparation set of Platelet Rich Plasma (PRP) of leucocytes is characterized in that,
the blood taking device comprises a rubber tourniquet (1), a blood taking needle (2), a first injector (3), a second injector (4), an anticoagulant storage device (5), a platelet-rich plasma extraction centrifuge tube (6) and a centrifuge (7), wherein the blood taking needle (2) is detachably connected with the first injector (3);
the platelet rich plasma extraction centrifuge tube (6) comprises: a centrifuge tube body (61) with a centrifuge tube cavity, a platelet leukocyte removing filter membrane arranged in the centrifuge tube cavity, a three-way pipe (8) arranged on the centrifuge tube body (61) and positioned below the leukocyte removing filter membrane, and a centrifuge tube cover (62) detachably connected at the tube orifice of the centrifuge tube body (61);
the tee (8) comprises: an upper longitudinal duct (81) and a lower longitudinal duct (82), and a transverse duct (83) connected between the upper longitudinal duct (81) and the lower longitudinal duct (82) and communicating with the upper longitudinal duct; the blood serum transfer pipeline (9), one end of the blood serum transfer pipeline (9) stretches into the centrifuge tube cavity for a certain length, the other end of the blood serum transfer pipeline (9) is connected with a port of the lower longitudinal pipeline (82), the upper end of the upper longitudinal pipeline (81) is communicated with the lower end of the centrifuge tube cavity, and the three-way pipe (8) is connected with a valve;
The centrifuge (7) comprises:
a host (71);
the rotating shaft (72) is rotatably arranged in the host machine (71) and driven by a driving motor in the host machine (71), and a plurality of rotors (78) are rotationally arranged in the host machine (71);
the first connecting rods (73) are in one-to-one correspondence with the rotors (78), one end of each first connecting rod (73) is fixedly connected with the corresponding rotating shaft (72), the other end of each first connecting rod (73) is provided with a fixed ring (74), and the rotors (78) are rotatably connected to the inner sides of the fixed rings (74) through connecting shafts (75) arranged front and back;
the outer centrifuge tubes (76) are provided with rotor (78) centrifuge cavities, and the platelet-rich plasma extraction centrifuge tubes (6) are arranged in the rotor (78) centrifuge cavities;
a rotor centrifuge chamber cover (77) detachably connected to the outer centrifuge tube (76) for closing or opening the rotor (78) centrifuge chamber;
the centrifuge (7) further comprises:
the pressure sensors are respectively arranged at the front side, the upper side and the left side of the contact part of the connecting shaft (75) and the rotor (78);
a first detecting device for detecting the rotation speed of the driving motor;
a second detection device for detecting the flow rate of whole blood in the platelet rich plasma extraction centrifuge tube (6);
The third detection device is used for detecting an included angle between the axis of the rotor (78) and the horizontal direction in the longitudinal rotation process of the rotor (78);
the controller is electrically connected with the pressure sensor, the first detection device, the second detection device and the third detection device, and the controller controls the driving motor to work with the controller based on the pressure sensor, the first detection device, the second detection device and the third detection device, and comprises the following steps:
step 1: the controller calculates the separation efficiency of the centrifugal machine (7) based on the second detection device, the third detection device and the formula (1);
Figure FDA0004179426250000021
wherein K isThe separation efficiency of the centrifuge (7) is natural constant, the value of e is 2.71828, rho is the density of whole blood, D is the diameter of a filter hole of a platelet-removing leukocyte filter membrane, D is the diameter of a platelet-rich plasma extraction centrifuge tube (6), mu is the dynamic viscosity of whole blood, sin is the sine alpha is the maximum included angle between the axis of the rotor and the horizontal direction in the longitudinal rotation process of the rotor, v is the flow velocity of the whole blood in the platelet-rich plasma extraction centrifuge tube, eta is the ratio of the volume of the whole blood in the platelet-rich plasma extraction centrifuge tube to the total volume in the platelet-rich plasma extraction centrifuge tube, A 1 For the current rotation speed of the driving motor of the centrifugal machine, A 3 The rated rotation speed of the driving motor of the centrifugal machine is set;
step 2: the controller calculates a safety evaluation coefficient Q of the centrifugal machine based on the step 1, the third detection device, the pressure sensor and the formula (2);
Figure FDA0004179426250000022
wherein A is 2 For the maximum safe rotation speed of the driving motor of the centrifugal machine, x is the average value of all the pressure sensor detection values at the front side, y is the average value of all the pressure sensor detection values at the upper side, z is the average value of all the pressure sensor detection values at the left side, and max (x, y, z) is the maximum value of the three values of x, y and z;
step 3: the controller compares the separation efficiency of the centrifugal machine with a preset separation efficiency reference value, and compares the safety evaluation coefficient of the centrifugal machine with a preset safety evaluation coefficient reference value;
when the separation efficiency of the centrifugal machine is greater than or equal to a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is greater than or equal to a preset safety evaluation coefficient reference value, the controller does not trigger an alarm device;
when the separation efficiency of the centrifugal machine is smaller than a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is smaller than a preset safety evaluation coefficient reference value, the controller controls the alarm device to send out a first alarm prompt;
When the separation efficiency of the centrifugal machine is greater than or equal to a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is smaller than the preset safety evaluation coefficient reference value, the controller controls the alarm device to send out a second alarm prompt, and the rotating speed of the driving motor of the centrifugal machine is reduced;
when the separation efficiency of the centrifugal machine is smaller than a preset separation efficiency reference value, and the safety evaluation coefficient of the centrifugal machine is larger than or equal to the preset safety evaluation coefficient reference value, the controller controls to increase the rotating speed of the centrifugal machine, and when the rotating speed is increased so that the separation efficiency of the centrifugal machine is equal to the preset separation efficiency reference value, the controller calculates the safety evaluation coefficient of the centrifugal machine again, if the safety evaluation coefficient of the centrifugal machine is larger than or equal to the preset safety evaluation coefficient reference value at the moment, the controller does not trigger an alarm device, and if the safety evaluation coefficient of the centrifugal machine is smaller than the preset safety evaluation coefficient reference value at the moment, the controller controls the alarm device to send a third alarm prompt.
2. A preparation kit for Platelet Rich Plasma (PRP) of white blood cells according to claim 1,
The first injector (3) is a 20ml injector and is used for collecting whole blood; the second injector (4) is a 10ml injector and is used for extracting bottom red blood cells, surface serum and PRP;
the anticoagulant stored in the anticoagulant storage device (5) is 4% sodium citrate anticoagulant.
3. A preparation kit for Platelet Rich Plasma (PRP) of white blood cells according to claim 2,
the centrifugal tube cover (62) is provided with a first connecting tube (63), an external thread is arranged outside the upper end of the first connecting tube (63), and a nut is connected to the external thread in a threaded manner and used for sealing the first connecting tube (63).
4. The leukocyte-depleted Platelet Rich Plasma (PRP) preparation kit of claim 1, wherein said leukocyte-removing filter is a concentrated platelet-removing polyester filter.
5. A preparation kit of a Platelet Rich Plasma (PRP) from white blood cells according to claim 3,
the using method of the leucocyte-rich platelet (PRP) preparation kit comprises the following steps:
step 1: obtaining whole blood, comprising:
step 11: 2ml of 4% sodium citrate anticoagulant was withdrawn using the 20ml syringe;
step 12: the rubber tourniquet (1) is tightly tied above the elbow joint, after the vein of the elbow is filled, the blood taking needle (2) is used for venipuncture, the tail part of the blood taking needle (2) is connected with a 20ml syringe after the step 11 is completed, and 18ml of whole blood is extracted;
Step 13: unscrewing a screw cap above the platelet and leukocyte removing filter membrane, connecting a 20ml syringe in the first connecting pipe (63) to filter the whole blood by the platelet and leukocyte removing filter membrane, and tilting the centrifuge tube for 30 degrees to enable the whole blood to flow to the bottom of the tube along the tube wall of the platelet-rich plasma extraction centrifuge tube (6);
step 2: blood centrifugation stratification, comprising:
placing the platelet-rich plasma extraction centrifuge tube (6) completed in the step 1 into a centrifugal cavity of a rotor (78) of a centrifuge (7), screwing a rotor centrifugal cavity cover (77), and adjusting centrifugal parameters to be: 1000g, 10min, 4 ℃; after centrifugation, the blood is layered, and comprises red blood cells at the bottom layer, platelet layers in the middle and serum at the uppermost layer;
step 3: obtaining platelet rich plasma, comprising:
step 31: rotating a valve of the three-way pipe (8), opening a transverse pipeline (83) of the three-way pipe (8), using the 10ml syringe to extract bottom red blood cells to the 2ml scale of the centrifuge tube, namely reserving 2ml bottom red blood cells, and closing the transverse pipeline (83);
step 32: then the valve is rotated, the lower longitudinal pipeline (82) of the tee pipe (8) is opened, and the 10ml injector is used for extracting the upper serum through the serum transfer pipeline (9);
step 33: the lateral channel (83) was again opened and concentrated platelet rich plasma was withdrawn using a 10ml syringe.
6. A preparation kit for Platelet Rich Plasma (PRP) of white blood cells according to claim 1,
still include belt cleaning device (10) for wash platelet rich plasma draws centrifuging tube (6), platelet removes white blood cell filter membrane joint platelet rich plasma and draws centrifuging tube (6) lateral wall, belt cleaning device (10) include:
the mounting seat (101), a plurality of mounting cavities (102) are arranged at the upper end of the mounting seat (101) at intervals and are used for mounting the platelet-rich plasma extraction centrifuge tube (6);
the fixing support (103) is fixedly connected to one side of the upper end of the mounting seat (101), and the upper end of the fixing support (103) is fixedly connected with a horizontal fixing plate (104);
the two fixing blocks (105) are fixedly connected to the lower end of the horizontal fixing plate (104) at left and right horizontal intervals;
the threaded rod (106) is horizontally arranged, two ends of the threaded rod (106) are rotationally connected with the two fixed blocks (105), a first thread and a second thread are respectively arranged on the outer walls of the left side and the right side of the center of the threaded rod (106), the rotation directions of the first thread and the second thread are opposite, and the threaded rod (106) is driven by a first rotating motor arranged on the fixed support (103) or the horizontal fixed plate (104);
The two sliding blocks (107), wherein the sliding blocks (107) are provided with horizontal threaded through holes, and the horizontal threaded through holes of the two sliding blocks (107) are respectively connected to the first threads and the second threads in a threaded manner;
a second mounting plate (108) disposed below the slider (107);
the upper ends of the two second connecting rods (109) are respectively and rotatably connected with the two sliding blocks (107), and the lower ends of the two second connecting rods (109) are respectively and rotatably connected to the second mounting plate (108);
the cleaning device comprises a plurality of first vertical fixing rods (110), wherein the upper ends of the first vertical fixing rods (110) are connected to a second mounting plate (108), liquid inlet cavities are formed in the first vertical fixing rods (110), and cleaning and clamping assemblies (111) are arranged on the first vertical fixing rods (110);
the cleaning and clamping assembly (111) comprises:
the arc plates (1111) are arranged at intervals on the periphery of the lower end of the vertical fixing rod, the upper ends of the arc plates (1111) are rotationally connected with the arc plates (1111), and the lower ends of the arc plates (1111) are provided with hook parts;
the brush hair is arranged on the outer side of the arc-shaped plate (1111), a connecting block (1112) is fixedly connected to the inner side of the arc-shaped plate (1111), and the inner side surface of the connecting block (1112) is a first conical surface;
the outer side of the pushing block (1113) is provided with a second conical surface, the outer side of the pushing block (1113) is in butt joint with the first conical surface, and the lower end of the pushing block (1113) is provided with a pushing ball (1114);
The second fixing rod (1115) is internally provided with a vertical flow channel, and the vertical flow channel is communicated with the liquid inlet cavity, and the upper end of the second fixing rod (1115) is fixedly connected to the lower end of the first vertical fixing rod (110);
the spring (1116) is sleeved outside the second fixing rod (1115), the upper end of the spring (1116) is fixedly connected with the lower end of the first vertical fixing rod (110), and the upper end of the spring (1116) is fixedly connected with the upper end of the pushing block (1113);
the miniature water pump is arranged on the mounting seat (101) or the fixing support (103) or the horizontal fixing plate (104), the water inlet end of the miniature water pump is connected with a water source, the water outlet end of the miniature water pump is connected with the liquid inlet cavity through a second connecting pipe, and an electromagnetic valve is arranged on the second connecting pipe;
a displacement sensor arranged on the horizontal fixing plate (104) for detecting the vertical displacement of the second mounting plate (108);
and the microcontroller is arranged on the mounting seat (101) or the fixing support (103) or the horizontal fixing plate (104) and is electrically connected with the miniature water pump, the displacement sensor and the first rotating motor.
7. A preparation kit for Platelet Rich Plasma (PRP) of white blood cells according to claim 6,
The upper end of the first vertical fixing rod (110) is rotationally connected to the second mounting plate (108), a second rotating motor for driving the first vertical fixing rod (110) to rotate is arranged on the second mounting plate (108), and the second connecting pipe is arranged at the center of the upper end of the first vertical fixing rod (110).
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